D-amino acid oxidase, and method for production of L-amino acid, 2-oxo acid, or cyclic imine
View Patent ↗The present invention relates to novel D-amino acid oxidase isolated and purified from Candida intermedia , a gene encoding the D-amino acid oxidase, a recombinant plasmid containing the gene, and a transformant into which the D-amino acid oxidase gene has been introduced, as well as a production method of D-amino acid oxidase including culturing the transformant. Moreover, the present invention relates to a production method of L-amino acids, 2-oxo acids or cyclic imines, which include reacting racemic amino acids with the D-amino acid oxidase, more preferably, a production method of L-amino acids, which includes reacting racemic amino acid with the D-amino acid oxidase, amino acid dehydrogenase and an enzyme having a coenzyme-regenerating activity. According to the present invention, L-amino acids, 2-oxo acids or cyclic imines can be produced with good efficiency in an industrial scale.
1. An isolated polynucleotide selected from (a)-(d):
(a) a polynucleotide comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 1;
(b) a polynucleotide comprising a polynucleotide sequence encoding a polypeptide having at least 90% amino acid identity to the amino acid sequence of SEQ ID NO: 1.
(c) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO: 2; and
(d) a polynucleotide comprising a polynucleotide sequence having at least 90% nucleic acid identity to the polynucleotide sequence of SEQ ID NO: 2,
wherein said polynucleotide sequence encodes a polypeptide with D-amino acid oxidase activity.
2. A recombinant plasmid comprising the isolated polynucleotide of claim 1 .
3. The recombinant plasmid of claim 2 , wherein said plasmid further comprises a second polynucleotide sequence encoding a protein with an activity different from that of D-amino acid oxidase.
4. The recombinant plasmid of claim 3 , wherein said second polynucleotide sequence encodes a polypeptide having an amino acid dehydrogenase activity or a coenzyme-regenerating activity.
5. An isolated transformant obtained by transforming a host microorganism with the recombinant plasmid of claim 2 .
6. The transformant of claim 5 , wherein said transformant further comprises a second recombinant plasmid encoding a polypeptide with an activity different from that of D-amino acid oxidase.
7. The transformant of claim 6 , wherein said second recombinant plasmid encodes a polypeptide having an amino acid dehydrogenase activity or a coenzyme-regenerating activity.
8. A method of producing D-amino acid oxidase, comprising culturing the isolated transformant of claim 5 , accumulating the polypeptide in the culture, and harvesting the polypeptide.